Structural characterization of the Sry 5’ regulatory region of bovine

By amplification of the polymerase chain reaction (PCR), the two fragments of 680 bp and 640 bp of 5’ regulatory region of Chinese bovine Sry gene were subcloned into pUC18. Sequence analysis shows that the fragment of total 1040 bp, which is highly conserved within bovine species, contains a start code ATG and a core promoter.     

牛ANGPTL1基因的电子克隆与序列分析

以牛的ANGPTL1基因为研究对象,利用生物信息学方法,对牛的ANGPTL1基因进行了电子克隆和序列分析,并对推导出的ANGPTL1蛋白结构与性质进行了初步分析。结果表明,牛的ANGPTL1基因序列长为2576bp,该基因的编码序列长为1476bp,编码492个氨基酸,编码序列的两翼有135bp的5’非编码区和785bp的3’非编码区。DNA序列的G＋C百分含量为44．31％,A＋T百分含量为55．69％。该基因的核苷酸序列与人、黑猩猩、鼠和狗ANGPTL1基因的cDNA序列的相似性分别为91％、90％、82％和93％。在氨基酸序列上与人、黑猩猩、鼠和狗的相似性分别为95％、95％、92％和95％。用氨基酸序列构建的进化树显示,在人、牛、黑猩猩、狗、褐鼠、原鸡几种动物中,牛与狗的亲缘关系最近。     

羚牛细胞色素b基因序列分析和系统进化研究

应用聚合酶链式反应（PCR）分别扩增了羚牛、绵羊、山羊、黄牛细胞色素b基因,并对其全序列（1140bp）进行了测定。其中羚牛细胞色素b基因序列属首次报道。通过对8种偶蹄类动物细胞色素b基因序列差异分析和基于序列差异所构建的分子系统树,发现羚牛与羊亚科的动物亲缘关系最近,与其他动物亲缘关系较远。表明将羚牛放入羊亚科较为合理,序列差异分析还表明羚羊约在距今500万年前（上新世）从牛类动物中分化出来,牛类分化时间约在600万年前的中新世（Miocene）。     

九龙牦牛CAST基因部分序列的克隆及其表达差异研究

根据牛钙蛋白酶抑制蛋白（Calpastatin, CAST） 设计并合成一对引物,从九龙牦牛肌肉组织提取总RNA,利用RT-PCR扩增获得九龙牦牛CAST基因部分片段,利用SQRT-PCR技术检测九龙牦牛各组织中CAST mRNA的表达差异．结果,成功克隆九龙牦牛CAST部分cDNA序列485bp,获得GenBank登录号为FJ483833;用DNAman软件分析发现九龙牦牛CAST基因与普通牛的核苷酸同源性依次为99％;SQRT-PCR组织表达检测表明,CAST基因在心脏、肝脏、脾脏、肺脏、肾脏、肌肉、脂肪中均表达,在心脏中表达最高,在脂肪组织中表达最低．     

Differentiation of Epinephelus moara from E. bruneus by improved nest-tetra-primer-specific PCR

Epinephelus moara and E. bruneus are closely related species in the genus Epinephelus (Perciformes, Serranidae). Their morphological similarity, changing color pattern at different stages and living conditions make them difficult to be differentiated. To identify these two species, an improved nest-tetra-primer-specific PCR assay was developed. Three specific molecular markers, the control region NC1 (394 bp), species-specific internal region ND2-M (268 bp) and ND2-B (122 bp), were identified in the mitochondrial ND2 gene from these two grouper species. Five markers were also discovered in the ITS1 regions of their nuclear ribosomal DNA, which were the control regions NC2 (588 bp) and NC3 (563 bp), and species-specific internal regions rDNA-M (426 bp), ITS1-M (488 bp) and ITS1-B (304 bp). This method provided a highly specific, precise, reliable and rapid molecular marker technique to discriminate between the two grouper species, as well as a new way of DNA identification to differentiate closely related species in fishes.     

Promoter activities of genes cpcT2 and cpcS2 in Nostoc PCC 7120

To study the promoter activities of genes cpcT 2 and cpcS 2,several upstream DNA fragments of these two genes with different lengths were selected.These fragments were fused with promoterless gfp(reporter gene) for constructing five recombinant plasmids.Then,these recombinant plasmids were transferred into Nostoc PCC 7120 by conjugation.The promoter activities of genes cpcT 2 and cpcS 2 were determined by observing the fluorescence of green fluorescent protein(GFP) with a fluorescence microscope.The result showed that the 1 300 bp(-1 300 to 0 bp) and 2 600 bp(-2 600 to 0 bp) upstream fragments of cpcT 2 had strong promoter activity and the promoter activity of the 680 bp(-680 to 0 bp) fragment was weaker than that of two above fragments of cpcT 2.The 2 000 bp(-2 000 to 0 bp) upstream fragment of cpcS 2 revealed weak promoter activity.This showed that a strong promoter of cpcT 2 was located in the upstream fragment between-680 and-1 300 bp.     

BMP－3及BMP－5在不同组织和细胞中的表达

报道ＢＭＰ－３及ＢＭＰ－５在不同组织细胞中的表达．将人的神经母细胞瘤ＳＫ细胞的总ＲＮＡ及人的脑、肝、胸腺、脾、胎盘及睾丸的总ＲＮＡ反转录成ｃＤＮＡ作为模板，利用设计的编码ＢＭＰ－３和ＢＭＰ－５成熟蛋白的专一性引物分别扩增出相应的片段．ＰＣＲ产物的琼脂糖凝胶电泳结果表明，ＢＭＰ－３及ＢＭＰ－５在不同组织细胞中表达类型不同．     

Effect of 5′-deletion ofArabidopsis profilin2 promoter on its vascular specific expression

Based on the published sequence of profilin2 promoter ofArabidopsis thaliana, a full-length promoter (1667 bp) was amplified by PCR. The 5′-end deletion fragments with length of 1380, 1153, 969 and 597 bp were then fused withgus (uidA) gene respectively. Constructed plant expression vectors were individually transferred intoKalanchoe laciniata and transgenic plants regenerated. GUS histochemical assay confirmed that the full-length promoter Pfn1.7 was vascular-specific. Deletion assays showed that profilin2 promoter could be divided into three parts. Deletion of fragment 1 (−1667—−1380 bp) resulted in constitutive expression, suggesting that element(s) responsible for vascular-specific expression might exist in this region. Fragment 2 located at −1153—−597 bp strongly inhibitedgus gene expression. Fragment 3 (−597—−1 bp) is considered as a basic domain of profilin2.